Скачать или смотреть From MEFs to Matrigel 3: Passaging hESCs from Matrigel onto Matrigel

Reference: https://app.jove.com/v/832/from-mefs-...
Passaging is a critical step in the cultivation and propagation of human embryonic stem cells (hESCs). It involves the transfer of hESCs from one layer of Matrigel, a specialized extracellular matrix, to another fresh layer of Matrigel. This process is necessary to provide the hESCs with a suitable environment for their continued growth and differentiation.

The first step in passaging is carefully detaching the hESCs from the initial Matrigel substrate. This is typically done using enzymatic digestion or mechanical methods to gently separate the cells from the matrix. It is crucial to ensure that the hESCs are detached without causing any damage or stress to the cells.

Once the hESCs are detached, they are then reseeded onto a fresh layer of Matrigel. This new layer provides a supportive and nourishing environment for the hESCs to adhere and proliferate. The Matrigel acts as a scaffold, mimicking the natural extracellular matrix found in the human body, and provides the necessary cues for the hESCs to maintain their pluripotency and viability.

Passaging is essential for maintaining the health and functionality of hESCs. As the cells continue to grow and divide, they can become overcrowded on the initial Matrigel layer, leading to reduced cell viability and compromised pluripotency. By transferring the hESCs onto a fresh layer of Matrigel, researchers can ensure that the cells have enough space and nutrients to continue their growth and differentiation.

Additionally, passaging allows researchers to expand the number of hESCs available for their studies and experiments. By carefully detaching and reseeding the cells, researchers can create multiple culture dishes or flasks containing hESCs, increasing the cell population for further analysis or manipulation.